Method for Identifying Human Growth Hormone Proteoform (hGHP) Pattern Biomarker

Abstract

The present disclosure provides a method for identifying a human growth hormone proteoform (hGHP) pattern biomarker. The method includes: collecting an hGH-secreting pituitary adenoma tissue sample and a normal pituitary tissue sample, and extracting tissue proteins, separately; conducting two-dimensional gel electrophoresis (2DGE), western blotting, and Coomassie brilliant blue (CBB) staining, and scanning visualized polyvinylidene fluoride (PVDF) membranes and 2D gels to obtain digital images; subjecting a corresponding protein in 2D gel spot to protein digestion with trypsin and purification, and conducting mass spectrometry identification and bioinformatics analysis to identify a GHP biomarker profile; and in combination with bioinformatics, using quantitative phosphoproteomics, quantitative ubiquitinomics, and quantitative acetylomics to identify post-translational modifications (PTMs) and splicing variations in GHP. The present disclosure can identify a change pattern of GHP between a GH-secreting pituitary adenoma tissue and a normal pituitary tissue. In total, 46 GHPs are identified in the GH-secreting pituitary adenoma tissue, and only 35 GHPs are identified in the normal pituitary tissue. Therefore, 11 GHPs are only present in the GH-secreting pituitary adenoma tissue.

Description

CROSS REFERENCE TO RELATED APPLICATION[0001]This patent application claims the benefit and priority of Chinese Patent Application No. 202110252781.3 filed on Mar. 9, 2021, the disclosure of which is incorporated by reference herein in its entirety as part of the present application.TECHNICAL FIELD[0002]The present disclosure belongs to the technical field of molecular biology, and specifically relates to a method for identifying a human growth hormone proteoform (hGHP) pattern biomarker.BACKGROUND ART[0003]Pituitary adenoma is one of the most-common primary intracranial tumors, with a prevalence rate of 17% worldwide. The abnormal growth hormone (GH) secretion occurring in pituitary adenoma can cause clinical diseases. In childhood, excessive GH secretion will cause abnormally-large bones and finally result in gigantism, but insufficient GH secretion will cause dwarfism. In adults, excessive GH secretion caused by pituitary adenoma will cause acromegaly. Pituitary adenoma causes som...

AI Technical Summary

Benefits of technology

The present disclosure identifies a biomarker expression profile and PTMs between GH-secreting pituitary adenoma tissue and normal pituitary tissue. This is used to search for a pattern biomarker abnormally expressed in GH-secreting pituitary adenomas and GH-related diseases. Different techniques, such as 2DGE, Western blotting with anti-GH antibodies, mass spectrometry, and bioinformatics, are used to identify a GHP pattern for the tissue. Additionally, quantitative phosphoproteomics, ubiquitinomics, and acetylomics are used to identify and quantify the phosphorylation, ubiquitination, and acetylation in hGHPs. The identified differences in GHPs and PTMs between the tumor and normal tissue can facilitate the early diagnosis, therapy, and prevention of GH-secreting pituitary adenomas and GH-related diseases.

Problems solved by technology

In childhood, excessive GH secretion will cause abnormally-large bones and finally result in gigantism, but insufficient GH secretion will cause dwarfism.

In adults, excessive GH secretion caused by pituitary adenoma will cause acromegaly.

Although there are various therapeutic methods such as drug therapy and surgical resection, a series of GH-related diseases caused by pituitary adenoma still cannot get satisfactory clinical treatment.

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